Bacterial adhesion to natural and man-made surfaces can be beneficial or detrimental, depending on the system at hand. Of vital importance is how the process of adhesion affects the bacterial metabolic activity. If activity is enhanced, this may help the cells colonize the surface, whereas if activity is reduced, it may inhibit colonization. Here, we report a study demonstrating that adhesion of both Escherichia coli and Bacillus brevis onto a glass surface resulted in enhanced metabolic activity, assessed through ATP measurements. Specifically, ATP levels were found to increase two to five times upon adhesion compared to ATP levels in corresponding planktonic cells. To explain this effect on ATP levels, we propose the hypothesis that bacteria can take advantage of a link between cellular bioenergetics (proton motive force and ATP formation) and the physiochemical charge regulation effect, which occurs as a surface containing ionizable functional groups (e.g., the bacterial cell surface) approaches another surface. As the bacterium approaches the surface, the charge regulation effect causes the charge and pH at the cell surface to vary as a function of separation distance. With negatively charged surfaces, this results in a decrease in pH at the cell surface, which enhances the proton motive force and ATP concentration. Calculations demonstrated that a change in pH across the cell membrane of only 0.2 to 0.5 units is sufficient to achieve the observed ATP increases. Similarly, the hypothesis indicates that positively charged surfaces will decrease metabolic activity, and results from studies of positively charged surfaces support this finding.Bacterial adhesion and biofilm formation are important to a wide array of fields, such as environmental, chemical, and biomedical engineering; food processing; materials science; marine science; and ecology and environmental sciences. A key consideration in the development of a biofilm is the initial interaction between the bacterium and the substrata and the way in which these interactions affect the metabolic activity of the cells. If the metabolic activity increases, this would help the cells colonize the surface, whereas if the metabolic activity decreases, then the cells may become inactive or die.There have been a number of studies that have examined the effects of attachment on bacterial metabolic activity. The first reported observation of this effect was made in 1943, where it was found that bacterial activity increased in the presence of glass surfaces, particularly with low nutrient concentrations (54). Since this first study, researchers have examined how various materials, including glass and polymer surfaces (10, 11, 24-26, 40, 44, 47), silicone surfaces (52), ceramic surfaces (32), dialysis membranes (18), activated carbon (7), clays (43), sands (31, 48), estuary particles (19), and ion exchange resins (46), affect the metabolic activity of attached bacteria.Different mechanisms have been examined in an attempt to explain how surfaces affect bacterial metabo...